title: Metabolic alterations in mammary cancer prevention by withaferin A in a clinically relevant mouse model.
abstract: BACKGROUND
  Efficacy of withaferin A (WA), an Ayurvedic medicine constituent, for prevention of mammary cancer and its associated mechanisms were investigated using mouse mammary tumor virus-neu (MMTV-neu) transgenic model.
  
  
  METHODS
  Incidence and burden of mammary cancer and pulmonary metastasis were scored in female MMTV-neu mice after 28 weeks of intraperitoneal administration with 100 µg WA (three times/week) (n = 32) or vehicle (n = 29). Mechanisms underlying mammary cancer prevention by WA were investigated by determination of tumor cell proliferation, apoptosis, metabolomics, and proteomics using plasma and/or tumor tissues. Spectrophotometric assays were performed to determine activities of complex III and complex IV. All statistical tests were two-sided.
  
  
  RESULTS
  WA administration resulted in a statistically significant decrease in macroscopic mammary tumor size, microscopic mammary tumor area, and the incidence of pulmonary metastasis. For example, the mean area of invasive cancer was lower by 95.14% in the WA treatment group compared with the control group (mean = 3.10 vs 63.77 mm2, respectively; difference = -60.67 mm2; 95% confidence interval = -122.50 to 1.13 mm2; P = .0536). Mammary cancer prevention by WA treatment was associated with increased apoptosis, inhibition of complex III activity, and reduced levels of glycolysis intermediates. Proteomics confirmed downregulation of many glycolysis-related proteins in the tumor of WA-treated mice compared with control, including M2-type pyruvate kinase, phospho glycerate kinase, and fructose-bisphosphate aldolase A isoform 2.
  
  
  CONCLUSIONS
  This study reveals suppression of glycolysis in WA-mediated mammary cancer prevention in a clinically relevant mouse model.
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
authors.forename: Eun-Ryeong
authors.initials: ER
authors.lastname: Hahm
authors.forename: Joomin
authors.initials: J
authors.lastname: Lee
authors.forename: Su-Hyeong
authors.initials: SH
authors.lastname: Kim
authors.forename: Anuradha
authors.initials: A
authors.lastname: Sehrawat
authors.forename: Julie A
authors.initials: JA
authors.lastname: Arlotti
authors.forename: Sruti S
authors.initials: SS
authors.lastname: Shiva
authors.forename: Rohit
authors.initials: R
authors.lastname: Bhargava
authors.forename: Shivendra V
authors.initials: SV
authors.lastname: Singh
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D016000:Cluster Analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006019:Glycolysis / Q000187:drug effects; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008175:Lung Neoplasms / Q000517:prevention & control* / Q000556:secondary; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism* / Q000517:prevention & control* / Q000821:virology; D008324:Mammary Tumor Virus, Mouse; D008515:Medicine, Ayurvedic; D051379:Mice; D008822:Mice, Transgenic; D011897:Random Allocation; D054358:Withanolides / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D054358:Withanolides; C009684:withaferin A